Treating method for suppressing hair growth

ABSTRACT

The present invention relates to a treating method for hair growth inhibition, which comprises administering (A) the extract of a plant of the family Juniperus or a malt. In addition, the present invention relates to a dermatologic composition for external use, which comprises (B) an elastase inhibitor or neutral endopeptidase inhibitor, and the above-described component (A) and/or (C) at least one proteolytic enzyme selected from the group consisting of papain, trypsin, chymotrypsin, pepsin, bromelain, ficin and pancreatin.

TECHNICAL FIELD

The present invention relates to a treating method and a dermatologiccomposition for external use, each for suppressing hair growth.

BACKGROUND ART

Although the body hair serves to biologically protect important organssuch as head, chest, hands and feet, the organ-protecting function ofthe hair is losing its importance with the appearance and development ofprotecting means such as clothes and protectors.

The hair was so far desired to be abundant. in recent years, however,the hair, particularly, that of hands and feet tends to be undesiredfrom the aesthetic viewpoint. To meet such a tendency, various body hairremoving methods have been developed and utilized. Specific examplesinclude a mechanical removing method using a shaver, hair-tweezers orthe like and, a method of rooting up the hair by using a depilatory anda method making use of the chemical action of a hair removing agent(EP-622069A1, WO97/44005).

These hair removing methods however give a physical or chemical stimulusto the skin. In addition, their effects do not last permanently, thoughthere is a little difference in durability among the methods, whichneeds the hair removing treatment again after the passage of apredetermined time. There is accordingly a demand or reducing thefrequency of the hair removing treatment.

An object of the present invention is therefore to provide a treatingmethod for hair growth inhibition which method is capable of effectivelysuppressing the growth of the body hair, thereby decreasing the removingfrequency of the body hair.

DISCLOSURE OF THE INVENTION

The present inventors have carried out an extensive investigation. As aresult, it has been found that a specific plant extract has excellenthair growth inhibitory effects and a dermatologic composition forexternal use which comprises it is useful as a hair growth inhibitor.

In one aspect of the present invention, there is thus provided atreating method for hair growth inhibition, which comprisesadministering an extract of the plant belonging to the family ofJuniperus or a malt extract.

In another aspect of the present invention, there is also provided adermatologic composition for external use, which comprises (A) anextract of the plant belonging to the family of Juniperus or a maltextract, (B) an elastase inhibitor or neutral endopeptidase inhibitor.

In a further aspect of the present invention, there is also provided adermatologic composition for external use, which comprises (A) anextract of the plant belonging to the family of Juniperus or a maltextract, (B) an elastase inhibitor or neutral endopeptidase inhibitor,and (C) at least one proteolytic enzyme selected from the groupconsisting of papain, trypsin, chymotrypsin, pepsin, bromelain, ficinand pancreatin.

In a still further aspect of the present invention, there is alsoprovided a dermatologic composition for external use, which comprises(B) an elastase inhibitor or neutral endopeptidase inhibitor, and (C) atleast one proteolytic enzyme selected from the group consisting ofpapain, trypsin, chymotrypsin, pepsin, bromelain, ficin and pancreatin.

In a still further aspect of the present invention, there is alsoprovided a treating method for hair growth inhibition, which comprisesadministering a composition containing the above-described components(A) and (B), a composition containing the above-descried components (A),(B) and (C), or a composition containing the above-described components(B) and (C).

BEST MODES FOR CARRYING OUT THE INVENTION

Examples of the plant belonging to the family of Juniperus and usable inthe present invention include Juniperus communis, Juniperus virginianaL., Juniperus morrisonicola Hayata and Juniperus formosana Hayata. Ofthese, Juniperus communis and Juniperus virginiana L. are particularlypreferred. These plants of the family of Juniperus are preferablyextracted at the leave or fruit portion thereof.

As a malt, that of wheat, barley, rye or oats is used.

The term “extract of a plant” as used herein means a solvent extractavailable by extracting the above-described portion of the plant in asolvent or through an extracting device such as Soxhlet extractor, orits dilute solution, concentrate or dry powder.

Examples of the solvent used for extraction include water, alcohols suchas methanol, ethanol, propanol and butanol, polyhydric alcohols such aspolypropylene glycol and butylene glycol, ketones such as acetone andmethyl ethyl ketone, esters such as methyl acetate and ethyl acetate,linear or cyclic ethers such as tetrahydrofuran and diethyl ether,halogenated hydrocarbons such as dichloromethane, chloroform and carbontetrachloride, hydrocarbons such as hexane, cyclohexane and petroleumether, aromatic hydrocarbons such as benzene and toluene, polyetherssuch as polyethylene glycol, and pyridines. They may be used eithersingly or in combination.

From the above-described extract, inactive impurities can be removed bya technique such as liquid-liquid partition or solvent-addingprecipitation. In the present invention, it is preferred to use such apurified extract. If necessary, the extract may be used afterdeodorization or decoloring in a known manner.

Although there is no particular limitation imposed on the form of thehair growth suppressing composition of the present invention, it ispreferred to formulate it as a hair-removing, depilatory or shavingcosmetic composition. Specific examples of such a cosmetic compositioninclude hair removers in the form of a paste, cream or aerosol,depilatories in the form of a wax, jelly or sheet, agents in the form ofa lotion or cream used for treatment after hair removal or depilation,pre-shaving agents such as pre-shaving lotion, shaving agents such asshaving cream, and after-shaving agents such as after-shaving lotion.

The above-described effective component is preferably contained in thehair growth inhibiting composition of the present invention in an amountof 0.00001 to 50 wt. %, particularly 0.0001 to 10 wt. %, as a dry solidscontent, from the viewpoints of the hair growth inhibitory effects andeconomy.

In the present invention, it is possible to use, together with thecomponent (A), an elastase inhibitor and/or neutral endopeptidaseinhibitor (B) and by using them in combination, the resultingdermatologic composition for external use has more improved hair growthinhibitory effects.

The “elastase” as used herein means one of proteolytic enzymes which arepresumed to play an important role for metabolism in the tissues of theliving body. It is known that the neutrophilic elastase closely relatesto the protection from infection or decomposition and regeneration ofthe damaged tissue, while the dermal-fibroblast-derived elastase takespart in the aging of the skin. The term “elastase inhibitor” as usedherein embraces respective substances having inhibitory activity againstthese elastases. Particularly preferred are substances inhibitingdermal-fibroblast-derived elastase, for example, those exhibitinginhibitory activity of at least 50% in the concentration of 1 mM in anenzyme activity measuring system which contains an enzyme solutionextracted from cultured human fibroblast with 0.1% Triton X-100/0.2Mtris-HCl buffer (pH 8.0) and, as a substrate,N-succinyl-Ala-Ala-Ala-p-nitroanilide.

Examples of such an elastase inhibitor include (1) phosphonic acidderivatives as described in Japanese Patent Application Laid-Open No.HEI 10-324611, (2) mercaptopropionic amide derivatives as described inJapanese Patent Application Laid-Open No. HEI 10-265360, and (3) plantsselected from ginger rhizome, hydrolyzed almond, birch, clove, rose hip,hawthorn, white birch and gambir or extracts, steam distillates orpressed products thereof.

The above-described phosphonic acid derivatives mean the phosphonic acidderivatives represented by the following formula (1):

(wherein, R¹ represents a hydrogen atom, a hydroxyl group, a substitutedor unsubstituted hydrocarbon group or a substituted or unsubstitutedsaccharide residue, R² represents a hydrogen atom, a substituted orunsubstituted hydrocarbon group or a substituted or unsubstitutedsaccharide residue, and R³ represents a hydrogen atom or—CH(R⁴)COOH (inwhich, R⁴ representing a hydrogen atom or a substituted or unsubstitutedhydrocarbon group), or salts thereof.

As the substituted or unsubstituted hydrocarbon group represented by R¹,R² or R⁴ in the above-described formula (1), either a saturatedhydrocarbon group or an unsaturated hydrocarbon group can be used andexamples include alkyl, alkenyl, alkynyl, cyclic alkyl, cyclic alkenyl,aromatic hydrocarbon and aralkyl groups. These hydrocarbon groups eachpreferably has 1 to 24 carbon atoms, particularly 1 to 18 carbon atoms.

Among the above-exemplified hydrocarbon groups, alkyl, cyclic alkyl,aromatic hydrocarbon and aralkyl groups are preferred. Preferredexamples of the alkyl group include linear or branched C₁₋₁₂ alkylgroups, with n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl andisoamyl groups being more preferred. Preferred examples of the cyclicalkyl group include 5- to 7-membered alicyclic alkyl groups, withcyclopentyl and cyclohexyl groups being more preferred. Preferredexamples of the aromatic hydrocarbon group include C₆₋₁₄ aromatichydrocarbon groups such as phenyl and naphthyl. Preferred examples ofthe aralkyl group include C₁₋₅ alkyl groups each substituted with aC₆₋₁₂ aromatic hydrocarbon group, such as 2-phenylethyl (=phenethyl),2-(1-naphthyl)ethyl and 2-(2-naphthyl)ethyl groups.

Examples of the group substitutable for these hydrocarbon groups includehalogen atoms, a hydroxy group, alkoxy groups, acyl groups, protected orunprotected amino groups and heterocyclic groups. Examples of thehalogen atom include chlorine, bromine and iodine. As the alkoxy groups,C₁₋₁₂ alkoxy groups such as methoxy, ethoxy and isopropoxy arepreferred. As the acyl groups, C₁₋₁₂ alkanoyl groups such as acetyl,propionyl and butyryl groups are preferred. Examples of the protected orunprotected amino groups include amino, acylamino, alkylamino anddialkylamino groups. As the heterocyclic group, preferred are 5- to14-membered monocylic or fused cyclic groups having 1 to 3 hetero atomssuch as nitrogen, oxygen and/or sulfur, for example, pyridyl,pyridazinyl, furyl, thienyl, indolyl, thiazolyl, imidazolyl, benzofuryland benzothienyl groups.

Examples of the saccharide residue include monosaccharide residue andoligosaccharide residue. Examples of the group substitutable for thesesaccharide residues include alkyl, acyl and aralkyl groups. As thealkyl, acyl and aralkyl groups, those exemplified above can bementioned.

These phosphonic acid derivatives can be prepared, for example, inaccordance with the process as described in Japanese Patent ApplicationLaid-Open No. HEI 5-105698.

The above-described mercaptopropionic amide derivatives meanmercaptopropionic amide derivative represented by the following formula(2):

(wherein, R⁵ represents a hydrogen atom or an acyl group, R⁶ representsa hydrogen atom or a substituted or unsubstituted hydrocarbon group, R⁷represents a hydrogen atom, a carboxyl group, an alkoxycarbonyl group, asubstituted or unsubstituted hydrocarbon group, a substituted orunsubstituted heterocyclic group or an acyl group and n stands for 1 to20) or salts thereof.

As the acyl group represented by R⁵ or R⁷ in the above-described formula(2), alkanoyl groups and arylcarbonyl groups can be mentioned. As thealkanoyl groups, C₁₋₁₂ alkanoyl groups such as acetyl, propionyl andbutyryl groups are preferred, while as the arylcarbonyl groups, C₇₋₁₅arylcarbonyl groups such as benzoyl, substituted benzoyl,naphthylcarbonyl and substituted naphthylcarbonyl groups are preferred.Examples of the group substitutable for the benzoyl or naphthylcarbonylgroup include alkyl, alkoxy, halogen, amino, hydroxy and alkanoyloxygroups.

As the substituted or unsubstituted hydrocarbon group represented by R⁶or R⁷, those exemplified above as R¹, R² or R⁴ can be mentioned.

As the heterocyclic group represented by R⁷, 5- to 14-memberedmonocyclic or fused ring groups each having 1 to 3 hetero atoms such asnitrogen, oxygen and/or sulfur are preferred. Specific examples includepyridyl, pyridazinyl, furyl, thienyl, indolyl, thiazolyl, imidazolyl,benzofuryl, benzothienyl, pyrrolidinyl, piperidinyl, morpholinyl andpiperazinyl groups. Examples of the group substitutable for theseheterocyclic groups include halogen atoms, hydroxyl group, alkoxylgroups, acyl groups and protected or unprotected amino groups. As thespecific examples of these substituents, those exemplified above as thesubstituent for the hydrocarbon group of R¹, R² or R⁴ can be mentioned.

Examples of the alkoxycarbonyl group represented by R⁷ include C₁₋₁₂alkoxycarbonyl groups, more specifically, methoxycarbonyl,ethoxycarbonyl, isopropoxycarbonyl and butoxycarbonyl groups.

These mercaptopropionic amide derivatives can be prepared, for example,in accordance with the process as described in Japanese PatentApplication Laid-Open No. SHO 57-24354.

The following are the typical compounds of the above-describedphosphonic acid derivatives and mercaptopropionic amide derivatives.

In the present invention, the ginger rhizome (Zingiberis Rhizoma) meansan underground stem of ginger (Zingiber officinale Roscoe) of the familyZingiberaceae; the hydrolyzed almond means a mixture available byhydrolyzing, in the presence of an acid or alkali, the seed (sweet seed)of almond (Prunus amygdalus Batsch) of the family Rosaceae, the clovemeans a bud of a clove (Syzygium aromaticum Merrill et Perry (Eugeniacaryophyllata Thunberg)), the rose hip means a fruit of a wild rose(Rosa multiflora Thunberg) of the family Rosaceae or plants analogousthereto, the hawthorn means a terrestrial part of a hawthorn (Craegusoxyacantha L.) of the family Rosaceae, the white birch means a leave,bark, or wood part of European birch (Betula alba L.) of the familyButulaceae, and the gambir means a dried and solidified aqueous extractprepared from the leaves or young twigs of Uncaria gambir Roxburgh ofthe family Rubiaceae. These plants have conventionally been used as acrude drug or food and are excellent in safety.

The term “plant extract” as used herein, except the hydrolyzed almond,means a solvent extract available by extracting the above-describedplant in the pulverized form with a solvent at room temperature or underheating, or extracting by an extractor such as Soxhlet extractor; or adiluted solution, concentrate or dry powder thereof.

In the present invention, it is possible to use plants as are or afterextraction, steam distillation or pressing. Examples of the solventusable for extraction include water, alcohols such as methanol, ethanol,propanol and butanol, polyhydric alcohols such as propylene glycol andbutylene glycol, ketones such as acetone and methyl ethyl ketone, esterssuch as methyl acetate and ethyl acetate, linear or cyclic ethers suchas tetrahydrofuran and diethyl ether, halogenated hydrocarbons such asdichloromethane, hydrocarbons such as hexane, cyclohexane and petroleumether, aromatic hydrocarbons such as toluene, polyethers such aspolyethylene glycol and pyridines. They may be used either singly or incombination.

On the other hand, the hydrolyzed almond can be obtained, for example,by adding 0.1 to 20 vol. % of an acid such as sulfuric acid,hydrochloric acid, acetic acid or phosphoric acid or 0.01 to 10N alkalisuch as sodium hydroxide or potassium hydroxide to a solvent such aswater, ethanol, propanol, butanol, propylene glycol or 1,3-butyleneglycol, preferably water or ethanol, dipping an almond in the resultingmixture usually at 3 to 100° C. and then removing an insoluble matter.The insoluble matter is preferably removed after adjustment of the pH toabout 7.0 with an alkali such as sodium hydroxide, potassium hydroxide,sodium carbonate or sodium bicarbonate, or an acid such as sulfuricacid, hydrochloric acid, acetic acid or phosphoric acid. As the solvent,a mixture of at least two solvents may be used.

The above-described extracts can be used after removing therefrominactive impurities by a technique such as liquid-liquid partition orsolvent-adding precipitation and if necessary, subjecting the residue todeodorization or decoloring in a known manner. Moreover, it is possibleto use the extracts as a more active fraction obtained by fractionationthrough suitable separating means such as gel filtration, chromatographyor precision distillation.

Neutral endopipetidase is an enzyme for decomposing opioid peptides suchas enkephalins or neuropeptides such as substance P and bradykinin.Although the inhibitor against this enzyme is known to be useful as asubstitute for a morphine type substance, that is, as an analgesic,there is no report suggesting the relation between the activity of thisenzyme and growth of the body hair.

As the neutral endopeptidase inhibitor to be used in the presentinvention, a dermal-fibroblast-derived neutral endopeptidase inhibitoris preferred. Examples of such an inhibitor include substances eachexhibiting at least 50 inhibitory activity in the concentration of 1 mMin an enzymatic activity measuring system obtained by adding, to an MESbuffer (100 mM, pH 6.5) added with sodium chloride (300 mM), an enzymesolution extracted from the cultured human fibroblast with a 0.1 TritonX-100/0.2 M tris-HCl buffer (pH 8.0) andglutaryl-Ala-Ala-Phe-4-methoxy-2-naphthylamine as a substrate.

Examples of the neutral endopeptidase inhibitor include the compoundsrepresented by the following formulas (a) to (g).

(a) Malonic amide derivatives represented by the formula (a) and saltsthereof:

[wherein, R⁸ represents a hydrogen atom, an alkyl group, an alkenylgroup or an aralkyl group;

-   -   R⁹ represents a hydrogen atom or a substituted or unsubstituted        alkyl, alkenyl or aralkyl group,    -   R¹⁰ represents a hydrogen atom, an alkyl group or an alkenyl        group, or may be coupled with R¹¹ to form a heterocycle with the        adjacent nitrogen atom,    -   R¹¹ represents a hydrogen atom, or a substituted or        unsubstituted alkyl, alkenyl or aralkyl group, or may be coupled        with R¹⁰ to form the above-described heterocycle,    -   R¹² represents a hydroxyl group, an alkoxy group, an alkenyloxy        group or an amino acid residue, and    -   p stands for an integer of 0 to 5].

(b) Hydroxamic acid derivatives represented by the formula (b) and saltsthereof (Japanese Patent Application Laid-Open No. Sho 58-77852).

(wherein, Ar¹ represents a substituted or unsubstituted phenyl group,R¹³ represents a hydrogen atom, an alkyl group or a methylthioalkylgroup, and Z represents a hydroxy, alkoxy, aralkyloxy, phenoxy, amino,alkylamino or dialkylamino group).

(c) Mercaptopropionylamide derivatives represented by the formula (c)and salts thereof (Japanese Patent Application Laid-Open No. Sho60-1366554).

(wherein, R¹⁴ represents a hydrogen atom or an acyl group, R¹⁵represents an aralkyl or heteroaralkyl group, R¹⁶ represents a hydrogenatom or an alkyl, aralkyl or heteroaralkyl group, R¹⁷ represents ahydroxy, alkoxy, aralkyloxy, heteroaralkyloxy, amino, alkylamino ordialkylamino group and m stands for 1 to 15).

(d) N-substituted butylamide derivatives represented by the formula (d)and salts thereof (Japanese Patent Application Laid-Open No. Sho61-502468).

(wherein, A and B each independently represents a hydroxymethyl,carboxyl, esterified carboxyl, carbamoyl or N-substituted carbamoylgroup, R¹⁸ and R¹⁹ each independently represents an alkyl, aryl,heteroaryl or aralkyl group and Y represents a substituted orunsubstituted alkylene group).

(e) Hydroxamic acid derivatives represented by the formula (e) and saltsthereof (Japanese Patent Application Laid-Open No. Sho 63-101353).

(wherein, R²⁰ represents a hydrogen atom or an acyl group, R²¹ and R²³each independently represents a hydrogen atom, an alkyl group or anaralkyl group, R²² represents CONH or NHCO, R²⁴ represents a hydrogenatom or an amino group and R²⁵ represents a hydroxy, alkoxy, amino orN-substituted amino group).

(f) Dipeptides represented by the formula (f):

[wherein, R²⁶ represents a hydrogen atom or an alkyl group or may becoupled with R²⁷ to form a heterocycle together with the adjacentnitrogen atom,

-   -   R²⁷ represents a hydrogen atom or a substituted or unsubstituted        alkyl or aralkyl group, or may be coupled with R²⁶ to form the        above-described heterocycle,    -   R²⁸ represents a hydrogen atom or an alkyl, alkenyl or aralkyl        group, and    -   1 stands for an integer of 0 to 4].

(g) Dipeptides represented by the formula (g):

(wherein, R²⁹ and R³⁰ each independently represents a hydrogen atom oran alkyl or aralkyl group).

In the above-described formulas (a) to (g), the alkyl part of the alkyl,alkoxy, alkyloxy or methylthioalkyl group preferably has 1 to 6 carbonatoms. As the aralkyl group, phenyl (C₁₋₆ alkyl) groups are preferred.As the heteroaralkyl group, heteroaryl(C₁₋₆ alkyl) groups are preferred,with pyridyl(C₁₋₆ alkyl), pyrimidinyl(C₁₋₆ alkyl) and purine(C₁₋₆ alkyl)groups being particularly preferred. As the alkylene group, those having1 to 8 carbon atoms are preferred.

As the aryl group, phenyl and naphthyl groups are preferred. As thearalkyloxy group, phenyl((C₁₋₆ alkyl) groups are preferred. As theheteroaralkyloxy group, heteroaryl(C₁₋₆ alkyl)oxy groups are preferred,with pyridyl(C₁₋₆ alkyl)oxy, pyrimidinyl(C₁₋₆ alkyl)oxy and purine(C₁₋₆alkyl)oxy groups being particularly preferred. As the alkenyl group,those having 2 to 6 carbon atoms are preferred. Examples of thesubstituent for the alkyl, aralkyl or phenyl group include carboxylgroup, halogen atoms and alkoxy groups. Examples of the heterocycleformed by R¹⁰ and R¹¹ or R²⁶ and R²⁷ include pyrrolidine and piperidinerings.

Among them, the malonic acid amide derivatives of the formula (a) anddipeptides of each of the formula (f) and (g) are particularlypreferred.

As the alkyl or alkenyl group in R⁸ or R¹⁰ of the formula (a), thosehaving 1 to 8 carbon atoms are preferred, of which those having 1 to 4carbon atoms are particularly preferred and alkyl groups having 1 to 4carbon atoms are more preferred. Among them, methyl, ethyl, n-propyl,isopropyl-, n-butyl, isobutyl and t-butyl groups are particularlypreferred. Examples of the aralkyl group include phenylalkyl,biphenylalkyl and naphthylalkyl groups, of which the phenyl(C₁₋₆ alkyl)groups, particularly benzyl group, are preferred.

As R⁸, a hydrogen atom, C₁₋₈ alkyl groups and phenyl(C₁₋₆ alkyl) groupsare particularly preferred.

As R¹⁰, a hydrogen atom is most preferred.

As R⁹ or R¹¹, a hydrogen atom, linear or branched C₁₋₁₂ alkyl or alkenylgroups and aralkyl groups are preferred, of which the hydrogen atom andlinear or branched C₃₋₆ alkyl or alkenyl groups are more preferred, andthe hydrogen atom, n-propyl group, isopropyl group, isobutyl group andtert-butyl group are particularly preferred. Examples of the aralkylgroup include phenylalkyl and naphthylalkyl groups, of which thephenyl(C₁₋₆ alkyl) groups are preferred, with the benzyl and phenetylgroups being more preferred.

Examples of the heterocycle formed by R¹⁰ and R¹¹ include pyrrolidineand piperidine rings, of which the pyrrolidine ring is preferred.

As the alkoxy or alkenyloxy group in R¹², alkoxy or alkenyloxy groupshaving 1 to 8, particularly 1 to 4 carbon atoms are preferred, with theC₁₋₄ alkoxy groups being more preferred. Examples of the amino acidresidue include essential amino acid residues. The amino group of thisamino acid forms an amide bond with the carbonyl group in the formula(1). As R¹², a hydroxyl group and C₁₋₄ alkoxy groups are preferred, withhydroxyl, methoxy, ethoxy, n-propyloxy, isopropyloxy, n-butyloxy,isobutyloxy and t-butyloxy groups being particularly preferred.

As p, 0 or 1 is most preferred.

In the formula (f), a hydrogen atom is preferred as R²⁶. As R²⁷, ahydrogen atom and methyl, isopropyl, isobutyl, t-butyl, benzyl,phenethyl, carboxymethyl and carboxyethyl groups are preferred. As R²⁸,a hydrogen atom and methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, t-butyl and benzyl groups are preferred. As 1, 0, 1 or 2 ispreferred.

In the formula (g), R²⁹ preferably represents a hydrogen atom, while R³⁰preferably represents a hydrogen atom or a methyl group.

Examples of the salts of the compounds of the formulas (a) to (g)include alkali metal salts, alkaline earth metal salts, amine salts,amino acid salts and acid addition salts. Preferred are alkali metalsalts and amino acid salts. The compounds of the formulas (a) to (g) mayhave optical activity. Their steric configuration may be any one of R, Sor racemic form, or they may be in a hydrated form.

Among the compounds represented by the formula (a), particularlypreferred are as follows:

The compound represented by the formula (a) can be synthesized, forexample, in accordance with the process of Nakano, et al. (Chem. Lett.,505-8(1990)) represented by the following reaction scheme:

(R⁸ to R¹² and p have the same meanings as described above).

The target compound is obtained by reacting the malonic acid half ester(h) with the amino acid ester (i) in the presence of a dehydratingcondensation agent and optionally reacting the resulting mixture with abase such as sodium hydroxide.

The compound represented by the formula (a) can also be synthesized inaccordance with the below-described process of Katsuki, et al. (Bull.Chem. Soc. Jpn., 49, 3207-3290(1976)).

(wherein R⁸ to R¹² and p have the same meanings as described above).

Described specifically, the target compound is obtained by reacting themalonic acid half ester acid chloride (j) with the amino acid ester (i)in the presence of a base and optionally alkylating the resultingmixture with an alkyl halide or hydrolyzing it with a base such assodium hydroxide.

Examples of the dipeptide represented by the formula (f) includePhe-Gly, Phe-β-Ala, Phe-Phe, Phe-Leu, Phe-Ala and Phe-Asp. They can besynthesized, for example, by the process as described in K. Ienaga, K.Higashihara and H. Kimura, Chem. Pharma. Bull., 35, 1249-1254(1987).Examples of the dipeptide represented by the formula (7) includeAsp-Phe-OMe (aspartame) and Asp-Phe. As the compound of the formula (f)or (g), commercially available one can also be used.

From the viewpoints of hair growth inhibitory effects and economy, thedermatologic external composition of the present invention usuallycontains the elastase inhibitor and/or neutral endopeptidase inhibitorin an amount of 0.00001 to 40 wt. %, particularly 0.0001 to 10 wt. %. Ifthis component is an extract or the like, the above-described amount isin terms of dry solid powder.

The dermatologic composition for external use according to the presentinvention, which contains the components (A) and (B), further contains(C) at least one proteolytic enzyme selected from papain, trypsin,chymotrypsin, pepsin, bromelain, ficin and pancreatin.

The dermatologic composition for external use, which contains thecomponent (B) and the above-described component (C), also has excellenthair growth inhibitory effects.

It is usually preferred to adjust the content of the proteolytic enzyme(component (C)) in the dermatologic external composition of the presentinvention to 0.00001 to 10 wt. %, particularly 0.0001 to 3 wt. % fromthe viewpoints of the hair growth inhibitory effects and economy.

The weight ratio of the component (B) to the component (C) in thecomposition of the present invention depends on the nature of them, butthe (B):(C) weight ratio preferably falls within a range of from 10:1 to1:20, particularly 1:1 to 1:2, from the viewpoint of the hair growthinhibitory effects.

The composition of the present invention can be prepared by mixing thecomponents (B) and (C) in a conventional manner but it is possible tomix them after enclose them in a liposome to heighten the percutaneousabsorption. Liposome can be prepared, for example, by a known mannerwith a phospholipid such as lecithin as a material (“Biomembrane”,written by Robert B Gennis, pp.74-77, Springer Verlag Tokyo Co., Ltd.(1990)).

To the composition for inhibiting hair growth or dermatologic externalcomposition according to the present invention, a keratolytic or acomponent having hair-growth controlling or depilatory action such asthioglycolic acid or a salt thereof can be added as needed, in additionto the above-described component. Examples of the keratolytic includelactic acid, bioprase, salicylic acid, glycolic acid, citric acid, malicacid, sulfur, resorcin, thioxolone, selenium disulfide and urea.Examples of the salt of thioglycolic acid include not only sodium salt,potassium salt, calcium salt and ammonium salt but also alkanolaminesalts such as monoethanolamine, diethanolamine and triethanolamine.Calcium thioglycolate is particularly preferred. Each of thesekeratolytics, or thioglycolic acid or salts thereof is preferably addedin an amount of 0.01 to 10 wt. %, particularly 0.05 to 5%.

In the composition of the present invention, various desired componentsusually employed for cosmetics, quasi-drugs or drugs are incorporatedwithin a range not impairing the effects of the present invention.Examples of such desired components include purified water, ethanol, oilcomponents, humectants, thickeners, antiseptics, emulsifiers, drugefficacy ingredients, powders, ultraviolet absorbers, colorants,perfumes and emulsion stabilizers. Use of the dermatologic externalcomposition according to the present invention as a cosmeticcomposition, particularly, hair-growth-inhibitory cosmetic compositionis preferred.

Examples of the oil component include liquid paraffin, vaseline,paraffin wax, squalane, bees wax, carnauba wax, olive oil, lanolin,higher alcohols, fatty acids, synthetic ester oils between a higheralcohol and fatty acid and silicone oil. Examples of the humectantinclude sorbitol, xylitol, glycerin, maltitol, propylene glycol,1,3-butylene glycol, 1,4-butylene glycol, sodium pyrrolidonecarboxylate,lactic acid, sodium lactate, polyoxypropylene fatty acid ester andpolyethylene glycol. Examples of the thickener include water-solublepolymers such as carboxyvinyl polymer, carboxymethyl cellulose,polyvinyl alcohol, carrageenan and gelatin, and electrolytes such assodium chloride and potassium chloride. Examples of the antisepticinclude urea, methyl paraben, ethyl paraben, propyl paraben, butylparaben and sodium benzoate. Those of the emulsifier include nonionicsurfactants such as polyoxyethylene alkyl ether, polyoxyethylene fattyacid ester, polyoxyethylene sorbitan fatty acid ester, glycerin fattyacid ester, polyglycerin fatty acid ester, polyoxyethylene glycerinfatty acid ester, polyoxyethylene hydrogenated castor oil andpolyoxyethylene sorbitol fatty acid ester. Illustrative of the powderinclude talc, sericite, mica, kaolin, silica, bentonite, vermiculite,zinc powder, mica, mica titanium, titanium oxide, magnesium oxide,zirconium oxide, barium sulfate, red oxide, iron oxide and ultramarine.

The compositions of the present invention have excellent hair growthinhibitory effects and have a high degree of safety for human body.

EXAMPLES

The present invention will hereinafter be described more specifically byExamples. It should however be borne in mind that the present inventionis not limited to or by these examples. The amount of a plant extract isindicated as a dry solids content.

Preparation Example 1

Preparation of a Juniperus Extract

The raw leaves of Juniperus virginiana L. (the Family Juniperus ofCupressaceae) of American growth were dipped in 45% ethanol for 3 to 5days for extraction. The supernatant was filtered to obtain the extractof Juniperus virginiana L. The resulting extract was found to have aspecific gravity as measured at 25° C. of 0.93±5% and a solidconcentration of 1.24%.

Preparation Example 2

Preparation of a Juniperus communis Extract

A raw fruit of Juniperus communis (the Family Juniperus of Cupressaceae)was dipped in 50% ethanol for 3 to 5 days for extraction. Thesupernatant was filtered to obtain the extract of Juniperus communis.The resulting extract was found to have a solid concentration of 1.5%.

Preparation Example 3

Preparation of a Malt Extract

The raw malt of Gramineae wheat (Triticum aestivum L.) of Americangrowth was dipped in 42% ethanol for 3 to 5 days. The supernatant wasfiltered to obtain the malt extract. The resulting extract was found tohave a specific gravity as measured at 25° C. of 0.94±5% and a solidconcentration of 2.71%.

Preparation Example 4

Preparation of a Ginger Rhizome Extract

A ginger rhizome was sliced. To 50 g of the sliced ginger rhizome, 500mL of 50% ethanol was added to dip the former in the latter, followed byfiltration, whereby the ginger rhizome extract solution was obtained.The resulting extract solution was concentrated. As a result, the solidcontent was found to be 2.59 g, meaning that the extract solution hadthe solid concentration of 0.52%.

Preparation Example 5

Preparation of a Hydrolyzed Almond Extract

The pulverized almond (50 g) was extracted under heating (60° C. for 2hours) with 200 mL of 0.1 mol/L sulfuric acid. The mixture was thenallowed to cool down and filtered, whereby the extract solution wasobtained. After neutralization of the resulting extract solution with0.2 mol/L sodium hydroxide, it was diluted with distilled water, whereby500 g of the hydrolyzed almond extract solution was obtained. Theevaporation residue was found to be 5.2 g.

Preparation Example 6

Preparation of a Burnet Extract

The burnet was sliced. To 50 g of the sliced burnet, 500 mL of water wasadded to dip the former in the latter, followed by filtration, wherebythe burner extract solution was obtained. As a result of concentration,the solid content was found to be 2.68 g, meaning that the extractsolution had a solid concentration of 0.54%.

Preparation Example 7

Preparation of a Clove Extract

The clove was sliced. To 50 g of the sliced clove, 500 mL of 95% ethanolwas added to dip the former in the latter, followed by filtration,whereby a clove extract solution was obtained. As a result ofconcentration, the solid content was found to be 1.46 g, meaning thatthe extract solution had a solid concentration of 0.29%.

Preparation Example 8

Preparation of a Rose Hip Extract

The rose hip was sliced. To 50 g of the sliced rose hip, 500 mL of waterwas added to dip the former in the latter, followed by filtration,whereby a rose hip extract solution was obtained. As a result ofconcentration, the solid content was found to be 2.28 g, meaning thatthe extract solution had a solid concentration of 0.46%.

Preparation Example 9

Preparation of a Hawthorn Extract

The hawthorn was sliced. To 50 g of the sliced hawthorn was added 500 mLof 50% ethanol to dip the former in the latter, followed by filtration,whereby a hawthorn extract solution was obtained. As a result ofconcentration, the solid content was found to be 3.09 g, meaning thatthe extract solution had a solid concentration of 0.62%.

Preparation Example 10

Preparation of a White Birch Extract

The white birch was sliced. To 50 g of the sliced birch was added 500 mLof 50 ethanol to dip the former in the latter, followed by filtration,whereby a white birch extract solution was obtained. As a result ofconcentration, the solid content was found to be 3.13 g, meaning thatthe extract solution had a solid concentration of 0.63%.

Preparation Example 11

Preparation of a Gambir Extract

The gambir was sliced. To 5 g of the sliced gambir was added 100 mL ofethanol to dip the former in the latter, followed by filtration, wherebya gambir extract solution was obtained. As a result of concentration,the solid content was found to be 2.51 g, meaning that the extractsolution had a solid concentration of 2.51%.

Synthesis Example 1

Synthesis of Compound 8

In 50 mL of chloroform were dissolved 16.05 g (114 mmol) of glycineethyl ester hydrochloride and 23.27 g (228 mmol) of triethylamine,followed by ice cooling to 5° C. Without changing the temperature, 10.00g (57 mmol) of ethylmalonic chloride was added dropwise to the resultingsolution. After completion of the dropwise addition, the disappearanceof the raw material was confirmed by thin layer chromatography and a 5%aqueous phosphoric acid solution was added to terminate the reaction.The organic layer was washed with distilled water and saturated saline,dried over anhydrous sodium sulfate and then distilled to remove thesolvent. The residue was subjected to column chromatography and elutedwith a mixed solvent of ethyl acetate and n-hexane. The solvent was thendistilled off, whereby 9.40 g (yield: 76%) of Compound 1 was obtained.

NMR (DMSO-d₆) δ: 2.48-2.51 (m, 6H), 3.30 (d, 2H, J=9 Hz), 3.85 (d, 2H,J=6 Hz), 4.02-4.20 (m, 4H), 8.49 (t, 1H, J=5 Hz).

Synthesis Example 2

Synthesis of Compound 9

In 50 mL of dehydrated tetrahydrofuran was dissolved 5.00 g (23.0 mmol)of Compound 1. The resulting solution was added to a suspension of 1.10g (27.6 mmol) of sodium hydride in 30 mL of tetrahydrofuran, followed byheating to 50° C. Benzyl bromide (3.74 g, 21.9 mmol) was slowly addeddropwise and at the same temperature, stirring was conducted for 3hours. Then, the reaction was terminated. After cooling, a 5% aqueousphosphoric acid solution was added and the resulting mixture wasextracted with 300 mL of ethyl acetate. The organic layer was washedwith a saturated aqueous solution of sodium bicarbonate and saturatedsaline, dried over anhydrous sodium sulfate and then distilled to removethe solvent. The residue was subjected to column chromatography andeluted with a mixed solvent of ethyl acetate and n-hexane. The solventwas then distilled off, whereby 3.89 g (yield: 55%) of Compound 2 wasobtained.

NMR (DMSO-d₆) δ: 1.16(t, 1H, J=7 Hz), 2.93-3.19(m, 2H), 3.71(t, 1H, J=7Hz), 3.80(d, 2H, j=6 Hz), 4.06(q, 4H, j=7 Hz), 7.13-7.29(m, 5H), 8.61(t,1H, J=6H).

Synthesis Example 3

Synthesis of Compound 10

In 30 mL of methanol was dissolved 3.00 g (9.8 mmol) of Compound 2,followed by the addition of a solution or 1.20 g (21.5 mmol) ofpotassium hydroxide (KOH) in 10 mL of water. The resulting mixture wasstirred at room temperature for 2 hours and then, the reaction wasterminated. After removal of methanol by distillation under reducedpressure, a 5% aqueous phosphoric acid solution was added to the residueand the crystals thus precipitated were collected by filtration. Theresulting crystals were washed with water and then dried under reducedpressure, whereby 2.17 g (yield: 88%) of Compound 2 was obtained.

NMR (DMSO-d₆) δ: 2.90(m, 2H), 3.59(t, 1H, J=7 Hz), 3.74(d, 2H, J=6 Hz),7.12-7.32(m, 5H), 8.37(t, 1H, J=6 Hz), 12.49(br.s, 2H).

Synthesis Examples 4 to 6

Compounds 11 to 13 were synthesized in a similar manner to SynthesisExamples 1 to 3 by using the amino acid ester and alkyl halide as shownin Table 1.

TABLE 1 Synthesis Synthesis Example process Amino acid ester Alkylhalide Yield NMR 4 2 — Bromo- 61% DMSO-d₆ 1.14-1.25(m, 6H), biphenyl-2.99-3.20(m, 2H), methane 3.77(t, 1H, J=8Hz), 3.87(d, 2H, J=8Hz),3.98-4.19(m, 4H), 7.24-7.65(m, 9H), 8.63(t, 1H, J=5Hz) 5 1,2,3 β-alaninemethyl Benzyl 55% DMSO-d₆ 2.15-2.39(m, 2H), ester bromide 2.90-3.09(m,2H), hydrochloride 3.10-3.34(m, 2H), 3.54(t, 1H, J=8Hz), 7.00-7.35(m,5H), 8.11(t, 1H, J=5Hz), 12.38(br.s, 2H) 6 1,3 L-phenylalanine — 68%DMSO-d₆ 2.84-3.10(m, 2H), methyl ester 3.17(s, 2H), hydrochloride4.42-4.53(m, 1H), 7.09-7.35(m, 5H), 8.35(d, 1H, J=8Hz), 12.24(br.s, 2H)Test 1Inhibition Test on Neutral Endopeptidase Activity of Cultivated HumanFibroblast

Normal human fibroblast commercially available from DainipponPharmaceutical Co., Ltd. was subcultured on a DME medium containing 10%bovine fatal serum and provided for the present test. The cells peeledby a rubber-policeman from a Petri dish were floated in aphosphate-buffered physiological saline, collected by a low-velocitycentrifuge, and washed three times with the same physiological saline.The cells were floated in a 0.1% Triton X-100/0.2M tris-HCl buffer (pH8.0), followed by ultrasonic pulverization, whereby an enzyme solutionwas obtained.

The enzyme activity was measured as follows: To 100 μL of an MES buffer(100 mM, pH 6.5) added with sodium chloride (300 mM), were added 2 μL ofthe enzyme solution, 1 μL of a solution of the invention compound and,as a substrate for the measurement of enzyme activity, 1 μL of 20 mMglutalyl-Ala-Ala-Phe-4-methoxy-2-naphthylamine, followed by reaction at37° C. for 1 hour. Phosphoramidon was added to the reaction mixture togive its final concentration of 0.4 μM and then, the reaction wasterminated. Aminopeptidase M having a final concentration of 20 mU wasadded to the reaction mixture and the resulting mixture were reacted at37° C. for 15 minutes. The amount of the resulting4-methoxy-2-naphthylamine was determined by measuring the fluorescentstrength at excited wavelength of 340 nm and fluorescent wavelength of425 nm by a fluorospectrophotometer and % inhibition of the compoundagainst enzyme activity was determined. Results are shown in Table 2.

Table 2 Test substance Concentration % inhibition Compound 8 1 mM 23.8 ±2.7 Compound 9 0.1 mM   74.5 ± 9.3 Compound 10 0.1 mM   91.2 ± 3.7Compound 11 0.1 mM   85.7 ± 2.2 Compound 12 0.1 mM   69.5 ± 4.3 Compound13 1 mM 49.3 ± 1.2 Phe-Gly 1 mM 49.4 Phe-β-Ala 0.1 mM   92.6 Phe-Phe 1mM 85.7 Phe-Leu 1 mM 72.9 Phe-Ala 1 mM 75.3 Phe-Asp 1 mM 85.4 Aspartame1 mM 30.2 Asp-Phe 1 mM 45.7

Example 1

Hair Growth Test (1) Using Mice

Each of the test substances was dissolved or suspended in an extractionsolvent to give a concentration as shown in Table 3, whereby thecorresponding hair growth inhibitor was prepared. The back of each of49-day-old C3H/HeNCrj mice (one group consisting of 20 mice) was shavedby 2×4 cm² by electric clippers and an electric shaver so as not toinjure its skin. From the next day, the inhibitor was applied by 20μL/once/day to the shaved portion for 4 weeks, while only the solventwas applied to a control group. In order to observe hair regrowth, thepicture of the shaved portion was taken at a fixed magnification and theday-dependent change of the area ratio of the regrowth hair (regrowthhair area/shaved area) was measured by an image analyzer.

As a result, apparent from Table 3, it has been found that thecomposition of the present invention exhibited excellent hair growthinhibitory action.

Test 3 Hair growth inhibition Concentration ratio (%) 3 weeks Testsubstance (dry solids content) after depilation Malt extract 0.136% 48.6Juniperus virginiana L. extract 0.062% 36.1 Juniperus communis extract0.075% 30.5 Burnet extract 0.270% 58.5 Malt extract + burnet extract0.136% + 0.270% 72.3 Juniperus virginiana L. extract + 0.062% + 0.270%63.9 burnet extract Malt extract + Juniperus virginiana 0.136% +0.062% + 0.270% 82.4 L. extract + burnet extract

Example 2

Hair Growth Test (2) Using Mice

(1) Preparation of a Test Sample

Component (B)

Each of Compounds 1 to 3 and 7 was dissolved in a 50% ethanol solution,whereby a 1 mM solution was prepared.

Compound (C)

Papain powder (produced by CALBIOCHEM-NOVABIOCHEM CORPORATION) wasdissolved in purified water to prepare a 2% aqueous solution, followedby the addition of an equal amount of ethanol to prepare a 1% ethanolsolution.

Trypsin powder (produced by Sigma Aldrich Corporation) was dissolved inHanks' Balanced Salt Solution (produced by Gibco BRL) having 1 mMEDTA-4Na dissolved therein, followed by the addition of an equal amountof ethanol to prepare a 1% ethanol solution.

Chymotrypsin powder (produced by Sigma Aldrich Corporation) was treatedin a manner similar to that employed for trypsin, whereby a 1% ethanolsolution was prepared.

The above-described components (B) and (C) were combined as shown inTable 4 as a test sample.

(2) Testing Method

The back of 6-week-old C3H mice (each group consisting of 5 mice) wasshaved by 2×4 cm² by electric hair clippers or an electric shaver so asnot to injure its skin. The sample was applied in an amount of 100μL/once to the shaved portion twice a day for 4 weeks. To a controlgroup, only a solvent was applied. Three weeks after depilation, thepicture of the shaved portion was taken at a fixed magnification inorder to observe the regrowth hair and the area of the hair regrowth(area of regrowth hair/shaved area) was compared with that of thecontrol group by using an image analyzer. The results are shown in Table4.

TABLE 4 Concen- Hair growth inhibition 3 weeks after depilation trationChymo- Test sample applied Solvent Papain 1% Trypsin 1% trypsin 1%Solvent 0 17.3 25.6 20.1 Compound 1 1 mM 58.7 77.5 84.7 80.2 Compound 21 mM 59.1 80.3 — — Compound 3 1 mM 64.8 83.6 88.5 — Compound 7 1 mM 59.777.9 — —

As apparent from Table 4, the test sample containing the elastaseinhibitor as the component (B) and proteolytic enzyme as the component(C) in combination exhibited excellent hair growth inhibiting effects.

Example 3

Hair Growth Inhibiting Lotion

(wt. %) (i) Polyoxyethylene hydrogenated castor oil  0.8 Ethanol 30.0(ii) Malt extract  0.1 (dry solids content) Sodium dodecyl sulfate  0.12Dodecyl methylamine oxide  0.18 Isopropyl alcohol 15.0 Benzyl alcohol15.0 Glycerin  2.0 Purified water Balance

Components belonging to (i) and those belonging to (ii) were dissolved,respectively. To (i) was added (ii), followed by uniform mixing understirring, whereby a hair growth inhibiting lotion was obtained.

Example 4

Hair Growth Inhibiting Cream

(wt. %) (i) Liquid paraffin 10.0 Squalane  7.0 Jojoba oil  3.0 Solidparaffin  3.0 Polyoxyethylene cetyl ether  2.0 Sorbitan sesquioleate 1.0 Potassium hydroxide  0.1 (ii) Juniperus virginiana L.  0.01 (drysolids content) Glycerin  3.0 Ethyl paraben  0.1 Purified water Balance

Components belonging to (i) and components belonging to (ii) weredissolved under heating, respectively. To (i) was added (ii), followedby uniform mixing under stirring. The resulting mixture was emulsifiedand cooled, whereby a hair growth inhibiting cream was obtained.

Example 5

Hair Growth Inhibiting Foam

(wt. %) (i) Malt extract  0.1 (dry solids content) Cetanol  0.1Propylene glycol  2.0 Dimethyl silicone oil  2.0 Polyoxyethylenehydrogenated castor oil  2.5 Liquid paraffin  1.0 Polyvinyl pyrrolidone 0.5 Methyl paraben  0.2 Ethanol 10.0 Purified water Balance (ii)Liquefied petroleum gas (propellant)  4.0

After uniform mixing of the components belonging to (i), the resultingmixture was filled in a container. The component (ii) was then filled inthe container in a conventional manner, whereby a hair growth inhibitingfoam was prepared.

Example 6

Aerosol

(wt. %) (i) Juniperus virginiana L. 0.01 (dry solids content) Cetanol1.2 Propylene glycol 4.0 Ethanol 8.0 Purified water balance (ii)Liquefied petroleum gas (propellant) 4.0

After uniform mixing of the components belonging to (i), the resultingmixture was filled in a container. The component (ii) was then filled inthe container in a conventional manner, whereby an aerosol was prepared.

Example 7

Hair Growth Inhibiting Lotion

The components belonging to (i) and the components belonging to (ii)were dissolved, respectively. To (i) was added (ii), followed by uniformmixing under stirring, whereby a hair growth inhibiting lotion wasobtained.

(wt. %) (i) Polyoxyethylene hydrogenated castor oil 0.8 Ethanol 30.0(ii) Compound 6 1.0 Papain 1.0 Lactic acid 2.0 Sodium dodecyl sulfate0.12 Dodecylmethylamine oxide 0.18 Isopropyl alcohol 15.0 Benzyl alcohol15.0 Glycerin 2.0 Purified water Balance

Example 8

Hair Growth Inhibiting Cream

The components belonging to (i) and the components belonging to (ii)were dissolved under heating, respectively. To (i) was added (ii),followed by uniform mixing under stirring. After emulsification, theemulsion was cooled, whereby a hair growth inhibiting cream wasobtained.

(wt. %) (i) Liquid paraffin 10.0 Squalane 7.0 Jojoba oil 3.0 Solidparaffin 3.0 Polyoxyethylene cetyl ether 2.0 Sorbitan sesquioleate 1.0Potassium hydroxide 0.1 (ii) Compound 7 1.0 Trypsin enclosed in liposome2.0 Glycolic acid 2.0 Calcium thioglycolate 0.5 Glycerin 3.0 Ethylparaben 0.1 Purified water Balance

Example 9

Hair Growth Inhibiting Foam

The components belonging to (i) were uniformly mixed and filled in acontainer. The container was then filled with (ii) in a conventionalmanner, whereby a hair growth inhibiting foam was prepared.

(wt.%) (i) Compound 5 1.0 Chymotrypsin 1.0 Salicylic acid 1.0 Calciumthioglycolate 0.5 Cetanol 0.1 Propylene glycol 2.0 Dimethyl silicone oil2.0 Polyoxyethylene hydrogenated castor oil 2.5 Liquid paraffin 1.0Polyvinyl pyrrolidone 0.5 Methyl paraben 0.2 Ethanol 10.0 Purified waterBalance (ii) Liquefied petroleum gas (propellant) 4.0

1. A method of inhibiting hair growth comprising administering to asurface of skin in need thereof an effective amount of (B) an elastaseinhibitor and (C) at least one proteolytic enzyme selected from thegroup consisting of papain, trypsin, chymotrypsin, pepsin, bromelain,ficin and pancreatin.
 2. The method of inhibiting hair growth of claim1, wherein said component (C) is papain.
 3. The method of inhibitinghair growth of claim 1, wherein said component (C) is trysin.
 4. Themethod of inhibiting hair growth of claim 1, wherein said component (C)is chymotrypsin.
 5. The method of inhibiting hair growth of claim 1,wherein said component (C) is pepsin.
 6. The method of inhibiting hairgrowth of claim 1, wherein said component (C) is bromelain.
 7. Themethod of inhibiting hair growth of claim 1, wherein said component (C)is ficin.
 8. The method of inhibiting hair growth of claim 1, whereinsaid component (C) is pancreatin.